Articles producció científica> Enginyeria Química

Low-Cost Platform for Multiplexed Electrochemical Melting Curve Analysis

  • Datos identificativos

    Identificador: imarina:9286517
    Handle: https://hdl.handle.net/20.500.11797/imarina9286517
    Autores:
    Chahin, NassifEscobar-Nassar, SantiagoOsma, JohannBashammakh, Abdulaziz S SAlYoubi, Abdulrahman O OOrtiz, MayreliO'Sullivan, Ciara K K
    Resumen:
    Detection and identification of single nucleotide polymorphisms (SNPs) have garnered increasing interest in the past decade, finding potential application in detection of antibiotic resistance, advanced forensic science, as well as clinical diagnostics and prognostics, moving toward the realization of personalized medicine. Many different techniques have been developed for genotyping SNPs, and ideally these techniques should be rapid, easy-to-use, cost-effective, flexible, scalable, easily automated, and requiring minimal end-user intervention. While high-resolution melting curve analysis has been widely used for the detection of SNPs, fluorescence detection does not meet many of the desired requirements, and electrochemical detection is an attractive alternative due to its high sensitivity, simplicity, cost-effectiveness, and compatibility with microfabrication. Herein, we describe the multiplexed electrochemical melting curve analysis of duplex surfaces tethered to electrodes of an array. In this approach, thiolated probes designed to hybridize to a DNA sequence containing the SNP to be interrogated are immobilized on gold electrodes. Asymmetric PCR using a ferrocene-labeled forward primer is used to generate this single-stranded redox-labeled PCR amplicon. Following hybridization with the probe immobilized on the electrode surface, the electrode array is exposed to a controlled ramping of temperature, with concomitant constant washing of the electrode array surface while simultaneously carrying out voltammetric measurements. The optimum position of the site complementary to the SNP site in the immobilized probe to achieve maximum differentiation in melting temperature between wild-type and single base mismatch, thus facilitating allelic discrimination, was determined

  • Otros:

    Autor según el artículo: Chahin, Nassif; Escobar-Nassar, Santiago; Osma, Johann; Bashammakh, Abdulaziz S S; AlYoubi, Abdulrahman O O; Ortiz, Mayreli; O'Sullivan, Ciara K K
    Departamento: Enginyeria Química
    Autor/es de la URV: Chahin, Nassif / O'SULLIVAN, CIARA KATHLEEN / Ortíz Rodríguez, Mayreli
    Palabras clave: Voltammetric measurements Snp detection Electrochemical melting curve analysis
    Resumen: Detection and identification of single nucleotide polymorphisms (SNPs) have garnered increasing interest in the past decade, finding potential application in detection of antibiotic resistance, advanced forensic science, as well as clinical diagnostics and prognostics, moving toward the realization of personalized medicine. Many different techniques have been developed for genotyping SNPs, and ideally these techniques should be rapid, easy-to-use, cost-effective, flexible, scalable, easily automated, and requiring minimal end-user intervention. While high-resolution melting curve analysis has been widely used for the detection of SNPs, fluorescence detection does not meet many of the desired requirements, and electrochemical detection is an attractive alternative due to its high sensitivity, simplicity, cost-effectiveness, and compatibility with microfabrication. Herein, we describe the multiplexed electrochemical melting curve analysis of duplex surfaces tethered to electrodes of an array. In this approach, thiolated probes designed to hybridize to a DNA sequence containing the SNP to be interrogated are immobilized on gold electrodes. Asymmetric PCR using a ferrocene-labeled forward primer is used to generate this single-stranded redox-labeled PCR amplicon. Following hybridization with the probe immobilized on the electrode surface, the electrode array is exposed to a controlled ramping of temperature, with concomitant constant washing of the electrode array surface while simultaneously carrying out voltammetric measurements. The optimum position of the site complementary to the SNP site in the immobilized probe to achieve maximum differentiation in melting temperature between wild-type and single base mismatch, thus facilitating allelic discrimination, was determined and applied to the detection of a cardiomyopathy associated SNP.
    Áreas temáticas: Spectroscopy Electrochemistry Chemistry, analytical Analytical chemistry
    Acceso a la licencia de uso: https://creativecommons.org/licenses/by/3.0/es/
    Direcció de correo del autor: mayreli.ortiz@urv.cat mayreli.ortiz@urv.cat
    Identificador del autor: 0000-0002-9423-0055 0000-0002-9423-0055
    Fecha de alta del registro: 2025-02-18
    Versión del articulo depositado: info:eu-repo/semantics/publishedVersion
    URL Documento de licencia: https://repositori.urv.cat/ca/proteccio-de-dades/
    Referencia al articulo segun fuente origial: Acs Measurement Science Au. 2 (2): 147-156
    Referencia de l'ítem segons les normes APA: Chahin, Nassif; Escobar-Nassar, Santiago; Osma, Johann; Bashammakh, Abdulaziz S S; AlYoubi, Abdulrahman O O; Ortiz, Mayreli; O'Sullivan, Ciara K K (2022). Low-Cost Platform for Multiplexed Electrochemical Melting Curve Analysis. Acs Measurement Science Au, 2(2), 147-156. DOI: 10.1021/acsmeasuresciau.1c00044
    Entidad: Universitat Rovira i Virgili
    Año de publicación de la revista: 2022
    Tipo de publicación: Journal Publications

  • Palabras clave:

    Analytical Chemistry,Chemistry, Analytical,Electrochemistry,Spectroscopy
    Voltammetric measurements
    Snp detection
    Electrochemical melting curve analysis
    Spectroscopy
    Electrochemistry
    Chemistry, analytical
    Analytical chemistry

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